Cloning And Analysis Of The Sequences Of NBS-LRR And STK From Wild Rice Species Based On The Transcripts

Wild rice contains a large number of favorable gene which have disappeared in the cultivated rice,especially the resistance genes to biotic stesses and abiotic stresses. Because the wild rice genetic background is complex, there are many problems and shortcomings in the use of traditional methods to clone them. The strategy of cloning by the candidates of Resistance gene is a new method to clone the resistance gene that developed in the last decade years, which using the conserved resistance gene sequence to gain resistance gene analogs and ultimately clone the new genes. We extracted total RNA from Wild rice at first, reverse transcript to obtain cDNA; then designed PCR primers by the NBS-LRR and STK kinase gene conserved domains and amplified cDNA of the wild rice.the expected size of the zone is obtained, and clone into the vector,then sequenced it,finally we obtained 11 NBS-LRR resistance gene sequence homology and 26 STK kinase resistance gene homologous. the NBS-LRR resistance gene homology sequences can be divided into three categories, and STK kinase resistance gene homologous sequences can be divided into 13 categories. using BLAST alignment to analyse the NBS-LRR sequences,we found all three types of NBS-LRR sequences have high homologous to the resistance gene RPR1 of rice, and mapped them preliminary into the region near the RPR1 in the chromosome 11 of the rice genome by the method of electronic mapping. we infered it is the result of replication and differentiation of the RPR1 gene family in the process of Evolutionary. in addition to a category of sequence could not find the high sequence similarity in the rice genome , STK kinase resistance gene analogs could be found more than 90% homologous to the rice genome, and 10 categories can be mapped to hypothetical gene. These sequences may provide as probes in the further gene cloning,develop to be molecular markers, or as an initial sequence of the silico cloning,which can provide support to the cloning of new resistance gene from wild rice.